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THE PLANT CELL, Vol 8, Issue 1 95-105, Copyright © 1996 by American Society of Plant Biologists
Genetic and Biochemical Dissection of Transgenic RNA-Mediated Virus Resistance
J. Goodwin, K. Chapman, S. Swaney, T. D. Parks, E. A. Wernsman and W. G. Dougherty
Department of Microbiology, Oregon State University, Corvallis, Oregon 97331-3804
RNA-mediated virus resistance has been observed in transgenic plants at
varying frequencies, suggesting that a nuclear requirement or other
pre-condition must be met. This study was undertaken to characterize
genetically transgenes that confer a highly resistant state to infection by
tobacco etch virus (TEV). Transgenic tobacco line 2RC-6.13, expressing an
untranslatable mRNA containing the TEV coat protein open reading frame, had
three distinct transgene integration events that segregated as two linkage
groups. A genetic series of plants that contained zero, one, two, or all
three transgene inserts in both homozygous and heterozygous conditions was
produced and examined. Genetic and biochemical data suggested that
RNA-mediated virus resistance is a multigenic trait in line 2RC-6.13; three
or more transgenes were necessary to establish the highly resistant state.
One or two transgene copies resulted in an inducible form of resistance
(i.e., recovery). Transcription rates and steady state RNA levels of the
transgene-derived transcript present in different members of the genetic
series supported a post-transcriptional RNA degradation process as the
underlying mechanism for transgene transcript reduction and virus
resistance. This degradation process appeared to initiate via cleavage of
specific sites within the target RNA sequence, as determined by RNA gel
blot and primer extension analyses of transgene-derived mRNA from various
transgenic plant lines.
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